LOCUS       Exported                7288 bp ds-DNA     circular SYN 22-OCT-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Oct 22, 2022 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7288
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     source          3141..3203
                     /organism="synthetic DNA construct"
                     /label=/dnas_title=pSmart-Kan-HC-Cascade
                     /note="/dnas_title=pSmart-Kan-HC-Cascade"
                     /note="color: #ffffff"
     source          4113..4181
                     /organism="synthetic DNA construct"
                     /label=/dnas_title=pSmart-Kan-HC-Cascade
                     /note="/dnas_title=pSmart-Kan-HC-Cascade"
                     /note="color: #ffffff"
     source          4836..4895
                     /organism="synthetic DNA construct"
                     /label=/dnas_title=pSmart-Kan-HC-Cascade
                     /note="/dnas_title=pSmart-Kan-HC-Cascade"
                     /note="color: #ffffff"
     promoter        197..1365
                     /label=EF1a P
                     /note="EF1 alpha promoter"
                     /note="color: #ffffff"
     CDS             1389..3140
                     /codon_start=1
                     /label=cas8c
                     /note="color: #993366"
                     /translation="ILHALTQYYQRKAESDGGIAQEGFENKEIPFIIVIDKQGNFIQLE
                     DTRELKVKKKVGRTFLVPKGLGRSGSKSYEVSNLLWDHYGYVLAYAGEKGQEQADKQHA
                     SFTAKVNELKQALPDDAGVTAVAAFLSSAEEKSKVMQAANWAECAKVKGCNLSFRLVDE
                     AVDLVCQSKAVREYVSQANQTQSDNVQKGICLVTGKAAPIARLHNAVKGVNAKPAPFAS
                     VNLSAFESYGKEQGFIFPVGEQAMFEYTTALNTLLASENRFRIGDVTAVCWGAKRTPLE
                     ESLASMINGGGKDKPDEHIDAVKTLYKSLYNGQYQKPDGKEKFYLLGLSPNSARIVVRF
                     WHETTVAALSESIAAWYDDLQMVRGENSPYPEYMPLPRLLGNLVLDGKMENLPSDLIAQ
                     ITDAALNNRVLPVSLLQAALRRNKAEQKITYGRASLLKAYINRAIRAGRLKNMKELTMG
                     LDRNRQDIGYVLGRLFAVLEKIQAEANPGLNATIADRYFGSASSTPIAVFGTLMRLLPH
                     HLNKLEFEGRAVQLQWEIRQILEHCQRFPNHLNLEQQGLFAIGYYHETQFLFTKDALKN
                     LFNEAKTA"
     CDS             3147..3200
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #993366"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3204..4052
                     /codon_start=1
                     /label=cas7
                     /note="color: #993366"
                     /translation="MTIEKRYDFVFLFDVQDGNPNGDPDAGNLPRIDPQTGEGLVTDVC
                     LKRKVRNFIQMTQNDEHHDIFIREKGILNNLIDEAHEQENVKGKEKGEKTEAARQYMCS
                     RYYDIRTFGAVMTTGKNAGQVRGPVQLTFSRSIDPIMTLEHSITRMAVTNEKDASETGD
                     NRTMGRKFTVPYGLYRCHGFISTHFAKQTGFSENDLELFWQALVNMFDHDHSAARGQMN
                     ARGLYVFEHSNNLGDAPADSLFKRIQVVKKDGVEVVRSFDDYLVSVDDKNLEETKLLRK
                     LG"
     CDS             4065..4091
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #993366"
                     /translation="YPYDVPDYA"
     CDS             4092..4112
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             4122..4178
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #993366"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4182..4823
                     /codon_start=1
                     /label=cas5
                     /note="color: #993366"
                     /translation="MRFILEISGDLACFTRSELKVERVSYPVITPSAARNILMAILWKP
                     AIRWKVLKIEILKPIQWTNIRRNEVGTKMSERSGSLYIEDNRQQRASMLLKDVAYRIHA
                     DFDMTSEAGESDNYVKFAEMFKRRAKKGQYFHQPYLGCREFPCDFRLLEKAEDGLPLED
                     ITQDFGFMLYDMDFSKSDPRDSNNAEPMFYQCKAVNGVITVPPADSEEVKR"
     CDS             4836..4895
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #993366"
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             4902..5282
                     /codon_start=1
                     /label=cas11
                     /note="color: #993366"
                     /translation="GLDRNRQDIGYVLGRLFAVLEKIQAEANPGLNATIADRYFGSASS
                     TPIAVFGTLMRLLPHHLNKLEFEGRAVQLQWEIRQILEHCQRFPNHLNLEQQGLFAIGY
                     YHETQFLFTKDALKNLFNEAKTA"
     polyA_signal    5286..5304
                     /label=BGH polyA
                     /note="BGH poly A signal (Bovine growth hormone)"
                     /note="color: #a6acb3"
     polyA_signal    5305..5491
                     /label=BGH polyA
                     /note="BGH poly A signal (Bovine growth hormone)"
                     /note="color: #a6acb3"
     misc_feature    5501..7288
                     /label=pSmart-Kan HC backbone
                     /note="pSmart-Kan HC backbone"
                     /note="color: #a6acb3"
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgga ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
      241 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc
      301 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
      361 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
      421 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg
      481 gcccttgcgt gccttgaatt acttccacct ggctccagta cgtgattctt gatcccgagc
      541 tggagccagg ggcgggcctt gcgctttagg agccccttcg cctcgtgctt gagttgaggc
      601 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
      661 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
      721 ctggcaagat agtcttgtaa atgcgggcca ggatctgcac actggtattt cggtttttgg
      781 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
      841 gcgagcgcgg ccaccgagaa tcggacgggg gtcggacggg ggtagtctca agctggccgg
      901 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
      961 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctccaggg
     1021 ggctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     1081 aaaggggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     1141 tccaggcacc tcgattagtt ctggagcttt tggagtacgt cgtctttagg ttggggggag
     1201 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1261 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1321 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaacaccg
     1381 ccaccatgat tctgcacgcc ctgacacagt actaccagcg gaaggccgaa agcgacggcg
     1441 gaattgccca agagggcttc gagaacaaag agatcccctt catcatcgtg atcgacaagc
     1501 agggcaactt catccagctc gaggacaccc gcgagctgaa agtgaagaaa aaagtgggcc
     1561 gcaccttcct ggtgcctaaa ggccttggca gaagcggcag caagagctac gaggtgtcca
     1621 acctgctgtg ggaccactac ggatacgtgc tggcctatgc cggcgagaag ggacaagaac
     1681 aggccgataa gcagcacgcc agcttcaccg ccaaagtgaa cgagctgaag caggccctgc
     1741 ctgatgatgc tggcgtgaca gctgtggccg cctttctgtc tagcgccgaa gagaagtcca
     1801 aagtgatgca ggccgccaac tgggccgagt gcgctaaagt gaagggctgc aacctgagct
     1861 tccggctggt ggatgaagcc gtggatctcg tgtgtcagtc taaggccgtg cgcgagtatg
     1921 tgtcccaggc caatcagacc cagagcgaca acgtgcagaa aggcatctgt ctggtcaccg
     1981 gcaaggccgc tcctattgcc agactgcaca atgccgtgaa gggcgtgaac gccaagcctg
     2041 ctcctttcgc ctctgtgaac ctgagcgcct ttgagagcta cggcaaagag cagggcttca
     2101 tcttccctgt gggagagcag gccatgttcg agtacaccac cgctctgaat accctgctgg
     2161 cctccgagaa cagattccgg atcggagatg tgaccgccgt gtgttgggga gccaagagaa
     2221 cacctctgga agagtccctg gccagcatga tcaatggcgg cggaaaggac aagcccgacg
     2281 agcacatcga cgccgtgaaa accctgtaca agagcctgta caacggccag taccagaagc
     2341 ctgacggaaa agagaagttc tacctgctgg gactgagccc caacagcgcc agaatcgttg
     2401 tgcggttctg gcacgagaca accgtggctg ccctgtctga gtctatcgcc gcttggtacg
     2461 acgacctgca gatggttcga ggcgagaaca gcccctatcc tgagtacatg cccctgccta
     2521 gactgctggg caacctggtg ctggacggca agatggaaaa cctgcctagc gacctgatcg
     2581 cccagatcac agatgctgcc ctgaacaaca gagtgctgcc tgtcagtctg ctgcaggcag
     2641 ccctgagaag aaacaaggcc gagcagaaga tcacctacgg cagagccagc ctgctgaagg
     2701 cctacatcaa ccgggccatc agagccggac ggctgaagaa catgaaggaa ctgaccatgg
     2761 gcctcgaccg gaacagacag gatatcggct atgtgctggg cagactgttc gccgtgctgg
     2821 aaaagattca ggccgaggcc aatcctggcc tgaacgccac aatcgccgac agatattttg
     2881 gcagcgccag cagcacacct atcgccgtgt ttggcaccct gatgagactg ctgcctcacc
     2941 acctgaacaa gctggaattc gagggcagag ccgtgcagct ccagtgggag atcagacaga
     3001 tcctggaaca ctgccagcgg ttccccaatc acctgaacct ggaacagcag ggactgtttg
     3061 ccatcggcta ctaccacgag acacagtttc tgttcaccaa ggacgccctg aagaacctgt
     3121 tcaacgaggc caagaccgcc ggctccgagg gccggggctc tctgctgacc tgtggcgacg
     3181 tggaggagaa ccccggcccc tccatgacca tcgagaagcg ctacgacttc gtgttcctgt
     3241 tcgacgtgca agacggcaac cccaacggcg atcctgatgc cggaaacctg cctagaatcg
     3301 accctcagac aggcgagggc ctcgtgacag atgtgtgcct gaagcggaaa gtgcggaact
     3361 tcatccagat gacccagaac gacgagcacc acgacatctt catcagagag aagggcatcc
     3421 tgaacaacct gatcgacgag gcccacgagc aagagaacgt gaagggcaaa gagaaaggcg
     3481 agaaaaccga ggccgccaga cagtacatgt gcagccggta ctacgacatc agaaccttcg
     3541 gcgccgtgat gaccaccggc aagaatgctg gacaagtgcg gggacctgtg cagctgacct
     3601 tcagcagatc catcgatccc atcatgaccc tggaacacag catcaccaga atggccgtga
     3661 ccaatgagaa ggacgccagc gaaaccggcg acaacagaac catgggcaga aagttcaccg
     3721 tgccttacgg cctgtaccgg tgccacggct ttatcagcac ccacttcgcc aagcagaccg
     3781 gcttcagcga gaacgacctg gaactgtttt ggcaggccct ggtcaacatg ttcgatcacg
     3841 atcactctgc cgccagaggc cagatgaatg ccagaggact gtacgtgttc gagcacagca
     3901 acaacctggg agatgcccct gccgacagcc tgttcaagag aatccaggtg gtcaagaaag
     3961 acggcgtgga agtcgtgcgg agcttcgacg attacctggt gtccgtggac gacaagaacc
     4021 tggaagagac aaagctgctg cggaagctcg gcggctctgt gggctatcct tacgacgtgc
     4081 cagactacgc ccctaagaaa aagcgcaaag tgggctccgg agccaccaac ttctccctgc
     4141 tgaagcaggc cggcgacgtg gaggagaacc ccggcccctc catgcggttc atcctggaaa
     4201 tcagcggcga cctggcctgc ttcacaagaa gcgagctgaa ggtcgagcgg gtgtcatacc
     4261 ctgtgatcac ccctagcgcc gccagaaaca tcctgatggc cattctgtgg aagcccgcca
     4321 tcagatggaa ggtgctgaag atcgagatcc tgaagcctat ccagtggacc aacatccggc
     4381 ggaacgaagt gggcaccaag atgagcgaga gaagcggcag cctgtacatc gaggacaaca
     4441 gacagcagcg ggcctccatg ctgctgaagg atgtggccta tagaatccac gccgacttcg
     4501 acatgacaag cgaggccggc gagagcgaca actacgtgaa gttcgccgag atgttcaagc
     4561 ggagagccaa gaagggccag tacttccacc agccttacct gggctgcaga gagttcccct
     4621 gcgacttcag actgctggaa aaggccgagg atggcctgcc tctggaagat atcacccagg
     4681 acttcggctt catgctgtac gacatggact tcagcaagag cgaccccaga gacagcaaca
     4741 acgccgagcc tatgttctac cagtgcaagg ccgtgaacgg cgtgatcact gtgcctccag
     4801 ccgatagcga ggaagtgaag agaggctctg tgggccagtg cacaaactac gccctgttga
     4861 agctggctgg agacgtggag tccaaccctg gaccctccat gggcctcgac cggaacagac
     4921 aggatatcgg ctatgtgctg ggcagactgt tcgccgtgct ggaaaagatt caggccgagg
     4981 ccaatcctgg cctgaacgcc acaatcgccg acagatattt tggcagcgcc agcagcacac
     5041 ctatcgccgt gtttggcacc ctgatgagac tgctgcctca ccacctgaac aagctggaat
     5101 tcgagggcag agccgtgcag ctccagtggg agatcagaca gatcctggaa cactgccagc
     5161 ggttccccaa tcacctgaac ctggaacagc agggactgtt tgccatcggc tactaccacg
     5221 agacacagtt tctgttcacc aaggacgccc tgaagaacct gttcaacgag gccaagaccg
     5281 cctgactcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     5341 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5401 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5461 gcaggacagc aagggggagg attgggaaga caatagcagg aacgaattca agcttgatat
     5521 cattcaggac gagcctcaga ctccagcgta actggactgc aatcaactca ctggctcacc
     5581 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     5641 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5701 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     6241 aaggtgagcc aggtgattac atttgggccc tcattagaaa aactcatcga gcatcaagtg
     6301 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     6361 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     6421 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     6481 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     6541 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     6601 cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     6661 gccgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     6721 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     6781 ccctgggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     6841 ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg accatctcat ctgtaacatc
     6901 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     6961 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     7021 taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat
     7081 atggctcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     7141 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     7201 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     7261 gtattgagcg atatctagag aattcgtc
//
